Atg3 Double Nickase Plasmid (h): sc-403308-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Atg3 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- Atg3 Double Nickase Plasmid (h) and Atg3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATG3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Atg3 Antibody (A-3): sc-393660
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Atg3 Double Nickase Plasmid (h) | sc-403308-NIC | 20 µg | $410.00 | |||
Atg3 Double Nickase Plasmid (h2) | sc-403308-NIC-2 | 20 µg | $410.00 |
ATG3 encodes Atg3, an E2-like enzyme essential for autophagy that catalyzes conjugation of LC3/ATG8 family proteins to phosphatidylethanolamine, enabling autophagosome membrane expansion and maturation. Through its role in the ATG12–ATG5–ATG16L1 ubiquitin-like conjugation cascade, ATG3 supports cytoplasmic quality control, organelle turnover, and adaptation to nutrient stress. Perturbation of ATG3-dependent autophagic flux can influence proteostasis, mitochondrial homeostasis, and innate immune signaling, processes frequently implicated in neurodegeneration, infection biology, and cancer cell stress responses. Accordingly, ATG3 is a useful node for studying selective autophagy, lysosome-dependent degradation, and cell survival programs under metabolic or proteotoxic stress.
Atg3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATG3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATG3. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt ATG3 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of ATG3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.